UNDERSTANDING THE PROBLEM
Stakeholder Interviews
Before investing our energies in doing wet lab and dry lab work, we wanted to verify that Vitamin A Deficiency is still a problem. We talked to people who work directly with vitamin A deficiency: nutritionists, public health specialists, anthropologists, and doctors. We created conversation about the problems with past attempts in vitamin A supplementation programs, what the supplementation process is like, the current status of vitamin A internationally and countries they are in, and the bottlenecks in the supplementation process.
Lourlin Ugdiman
Government Public Health & Nutrition Worker
Lourlin is a Public Health & Nutrition Worker working for the government in Arayat, Philippines. It was crucial for us to understand the status of Vitamin A deficiency in the Philippines, which was once a hotspot for VAD-caused child blindness. Lourlin also provided us insights on how supplements from the government reach the community.
Vitamin A is supplied every 6 months in the community every 6 months. The government provides the supplements while healthcare and nutrition workers like Lourlin provide them in schools, municipal centers, clinics, and individual houses that have difficulty going to these centers. The supplements they provide are sourced externally, such as India and China.
“Nagbibigay kami sa community ng vitamin A every 6 months. Ang pagbibigay namin ay tuwing April at October. Ayan (letrato) ang bigay ng Rural Health Unit sa DOH (Department of Health). Yung 200,000 IU binibigay namin sa mga batang 12 months to 59 months. Ang 100,000 IU ay kulay blue ay pang 9 months to 11 months. Hindi na katulad ng dati kasi now nasusuplayan ng government yung kailangan ng mga bata nga ng mga suppliment. Pero di nagiging sapat kasi sa kakulangan ng pagkain sa kani-kanilang pamilya. Kung may budget, gamit, at may personnel, para sa pag-gawa ng vitamin A, bakit hindi? Willing makipagtulungan ang gobyerno para sa ikauunlad at kapakanan ng kalusugan ng mga tao sa kumunidad.”
In our interview, Lourlin says “We supply vitamin A every 6 months at April and October. That (picture) is given by the Rural Health Unit in the Department of Health. The 200,000 IU we give to children 12-59 months, while the 100,000 IU which is blue (bottle) we give to kids 9 to 11 months. It’s not (VAD cases) like before because now the government provides the supplements. (As for the bioreactor) If there are finances, resources, and personnel, why not? The government is willing to work together as long as it’s for the development and welfare of the people and community’s health.”
Dr. Charles Mather
Information here
Dr. Charles Mather is a professor of social and cultural anthropology, sub-Saharan anthropology, and medical anthropology at the University of Calgary. As an individual with considerable experience in sub-Saharan cultures and medical systems, we contacted Dr. Mather for insight into how sub-Saharan regions of Africa are affected by Vitamin A deficiency, and how they may go about implementing our solution.
unicorn Need to change this
Dr. Sanou Dia
Information here
Dr. Sanou is a public health nutritionist who has worked with UNICEF, National Public Health Laboratory, and National Nutrition Center in Burkina Faso. He received a Ph.D. in Nutrition from Laval University and completed his postdoctoral training in aboriginal health at the University of Alberta. A graduate from the University of Ouagadougou (B.Sc. and M.Sc.), Professor Sanou also holds a Certificate in information technology and nutrition from the Swedish University of Agricultural Sciences.
The goal of our meeting to understand what bigger organizations such as UNICEF do to fight VAD. We also wanted to talk to him more specifically about the steps we would need to take from creating a product in the lab to its deployment in communities. He gave us insights about the vitamin A fiasco and the reasons why projects such as golden rice and other biofortified crops don’t work. This includes farmers preferring earning money for planting exportable crops over biofortified ones. Additionally, carotenoid to retinol bioconversion in the human body is minimal since the crops do not have a steady supply and take long to grow. With biofortification still being a new technology and thus making a very small part of the diet, Dr. Sanou says that having a regular supply of vitamin A rather than twice a year would be “much better”.
He works with communities to have “micronutrient week” or “vitamin A week” where healthcare workers mass supply vitamin A and educate them about its importance in diet. His concern though, is that even with more supplements sufficient for more than 100% of the country, there will still be individuals who will never access them due to issues such as distance and lack of proper roads and infrastructures.
We learned that universal supplementation of vitamin A for children under 2 years of age is coupled with deworming. This is because intestinal parasites perforate the intestines and prevent the maximal absorption of the vitamin.
Dr. Sanou explained that moving to a larger scale for supplementation, we would need a technologically feasible and impactful project. After obtaining replicates of our results, he advised us to reach out to funding institutions Canadian Institutes of Health Research and produce scientifically sound data. Then, we would work on the implementation and scaling up of the project before reaching out to humanitarian organizations and larger international funding institutes for distribution of our product.
Dr. Warren Wilson
Information Here
Dharamwati
Information Here
Dharamwati is an Accredited Social Health Activist (ASHA) worker in the Health Department of Delhi serving farming communities near Delhi. As an ASHA worker, she visits houses and conducts surveys about the health and economic status of the household. The ASHA workers in coordination with Aanganwadi workers, enroll the children and keep regular tab on their weight, physical and mental growth by providing them with nutritious food and involving them in various constructive and educative activities.
We learned that they perform diagnostic tests of the children done, in order to identify and monitor vitamin, calcium and iron deficiencies. They also help enroll expecting women in Aanganwadis, so that proper food and economic help (Government Sponsored) can be provided to them.
Dharamwati said that 20-25% of children in her area suffer from Vitamin A deficiency. Together with ASHA and the government, they provide Vitamin A drops every six months to children from 9 months to 5 years of age. They also counsel mothers to incorporate vitamin A rich foods in their diet.
She brought up her concern of the non-availability of the Vitamin A syrup sometimes, and community members resisting administration of the vitamin to their children. This inspired us to develop a sustainable synthetic biology solution to vitamin A deficiency by using plentiful resources such as agricultural waste, that would normally be discarded.
What did we discover?
Current State of Vitamin A Deficiency
Our HP contacts who work in the Philippines, Ghana, India, and Burkina Faso told us that VAD still affects a significant percentage of the children in their countries. The government plays a big role in promotion of growing beta-carotene biofortified crops and supplying vitamin A (retinol) capsules. However, these efforts are not enough.
Ineffective Delivery
Past approaches such as the Golden Rice project and biofortification of other staple crops through genetic engineering did not work well because plants need months to grow and farmers would suffer economic losses because the biofortified crops can contaminate their non-fortified crops and prevent its export. Additionally, vitamin A supplementation through gel capsules is only done biannually. Health and nutritional organizations allot for more than 100% coverage of supplements, but social workers and mobile nurses are not always able to reach rural areas.
Lack of Sustainability
Many institutions from western companies have provided VAD-vulnerable countries with resources to combat the problem, but we learned that they don’t work well because these institutions do not leave infrastructure for the people to work with. As a result, their efforts go in vain as soon as they leave the country and stop providing resources. Many of these efforts also do not consider the culture and available resources in the country they are helping. Lastly, the money has to come from someone’s pocket. Even with cheap vitamin A capsule prices, transportation and distribution costs can still be economically straining. Most importantly, the people we are working with do not want external aid enforced on them. They want something sustainable-- something THEY can have ownership and accountability of.
IDEATE
Propose Solutions
Having identified key problems in the vitamin A supplementation distribution, our team started brainstorming ideas. Upon careful consideration of our stakeholders’ needs, we decided on using a chassis producing β-carotene targeted for consumption like nutritional yeast. Upon conducting literature review, we decided on working with Rhodotorula toruloides, a red oleaginous yeast which naturally produces β-carotene. Then, we would genetically modify it with a series of cellulases to use agricultural waste as a feedstock. For added nutritional benefits, we will engineer two additional genes to our chassis for expression of EPA, an omega-3 fatty acid known to have cognitive benefits. A light-inducible kill switch would also be added as a biocontainment measure. For added safety features and optimized growth conditions, our yeast would be grown in a laboratory bioreactor. With these attributes, the yeast could be grown in a community center, clinics, or dispensaries-- improving vitamin coverage and delivery in a sustainable manner.
To learn more about these solutions, please click the buttons below.
Verifying Need
With a proposal on hand, we talked to our end users to verify its need and discuss its implementation in their workplace and communities. We talked to healthcare workers and nutritionists in rural areas in South Asia, Sub-Saharan Africa, and Southeast Asia which are among the most VAD-vulnerable areas.
Lourlin Ugdiman
Public Health & Nutrition
Lourlin is a Public Health & Nutrition Worker working for the government in Arayat, Philippines. It was crucial for us to understand the status of Vitamin A deficiency in the Philippines, which was once a hotspot for VAD-caused child blindness. Lourlin also provided us insights on how supplements from the government reach the community.
Vitamin A is supplied every 6 months in the community every 6 months. The government provides the supplements while healthcare and nutrition workers like Lourlin provide them in schools, municipal centers, clinics, and individual houses that have difficulty going to these centers. The supplements they provide are sourced externally, such as India and China.
“Nagbibigay kami sa community ng vitamin A every 6 months. Ang pagbibigay namin ay tuwing April at October. Ayan (letrato) ang bigay ng Rural Health Unit sa DOH (Department of Health). Yung 200,000 IU binibigay namin sa mga batang 12 months to 59 months. Ang 100,000 IU ay kulay blue ay pang 9 months to 11 months. Hindi na katulad ng dati kasi now nasusuplayan ng government yung kailangan ng mga bata nga ng mga suppliment. Pero di nagiging sapat kasi sa kakulangan ng pagkain sa kani-kanilang pamilya. Kung may budget, gamit, at may personnel, para sa pag-gawa ng vitamin A, bakit hindi? Willing makipagtulungan ang gobyerno para sa ikauunlad at kapakanan ng kalusugan ng mga tao sa kumunidad.”
In our interview, Lourlin says “We supply vitamin A every 6 months at April and October. That (picture) is given by the Rural Health Unit in the Department of Health. The 200,000 IU we give to children 12-59 months, while the 100,000 IU which is blue (bottle) we give to kids 9 to 11 months. It’s not (VAD cases) like before because now the government provides the supplements. (As for the bioreactor) If there are finances, resources, and personnel, why not? The government is willing to work together as long as it’s for the development and welfare of the people and community’s health.”
Dr. Charles Mather
Dry Lab
Dr. Charles Mather is a professor of social and cultural anthropology, sub-Saharan anthropology, and medical anthropology at the University of Calgary. As an individual with considerable experience in sub-Saharan cultures and medical systems, we contacted Dr. Mather for insight into how sub-Saharan regions of Africa are affected by Vitamin A deficiency, and how they may go about implementing our solution.
After expressing his support of and enthusiasm for our idea, Dr. Mather provided us with valuable feedback that will no doubt inform our design process. For instance, he informed us that some rural areas lack access to electricity, which poses a challenge to the design of our bioreactor, into which we were considering integrating an electric air pump. He cited solar-powered air pumps as a feasible alternative, while also suggesting that the culturing process be modified to incorporate stirring, rather than needing an air pump. As extreme heat decreases the efficiency of the yeast, Dr. Mather also cautioned us of the climate in Ghana, which can reach over 50°C in the dry season.
Furthermore, Dr. Mather informed us that people in Northern Ghana are very willing to take initiative with regards to technologies that will have a positive impact, suggesting that a representative travel to the community to educate people about how to culture, harvest, and cook the yeast, as well as the health benefits of the yeast. Considering the fact that Northern Ghana has an active beer brewing industry, where local women take turns brewing beer in an outdoor setup for the entire community. Dr. Mather recommended adapting the yeast culturing and harvesting process so that it can be integrated into existing beer brewing infrastructure. He emphasized that in many cases it would not be important to tell the people exactly how to assemble or culture the yeast. Instead, in many respects it might be preferable to simply tell them the growing needs of the yeast and allow them to decide how best to meet those needs.
Dr. Mather also provided some insight into the cultural associations surrounding blindness, stating that spiritual healing medicines and shrines play a big role in Ghanaian culture. However, he cited the Ghanaian people as highly pluralistic and open-minded, stating that they are willing to adopt new beneficial technologies and medicines as they become available. In order to implement our solution in a long-term and sustainable manner, he recommends beginning at the village level. Once our technology has been adopted by the village, he then recommends expanding into surrounding areas and moving up the chain of command.
Dr. Sanou is a public health nutritionist who has worked with UNICEF, National Public Health Laboratory, and National Nutrition Center in Burkina Faso.
He received a Ph.D. in Nutrition from Laval University and completed his postdoctoral training in aboriginal health at the University of Alberta.
A graduate from the University of Ouagadougou (B.Sc. and M.Sc.), Professor Sanou also holds a Certificate in information technology and nutrition from
the Swedish University of Agricultural Sciences.
The goal of our meeting to understand what bigger organizations such as UNICEF do to fight VAD.
We also wanted to talk to him more specifically about the steps we would need to take from creating a product in the lab to its deployment in communities.
He gave us insights about the vitamin A fiasco and the reasons why projects such as golden rice and other biofortified crops don’t work.
This includes farmers preferring earning money for planting exportable crops over biofortified ones.
Additionally, carotenoid to retinol bioconversion in the human body is minimal since the crops do not have a steady supply and take long to grow.
With biofortification still being a new technology and thus making a very small part of the diet, Dr. Sanou says that having a regular supply of vitamin A
rather than twice a year would be “much better”.
He works with communities to have “micronutrient week” or “vitamin A week” where healthcare workers mass supply vitamin A and educate them about its importance in diet.
His concern though, is that even with more supplements sufficient for more than 100% of the country, there will still be individuals who will never access them due to
issues such as distance and lack of proper roads and infrastructures.
Dr. Sanou explained that moving to a larger scale for supplementation, we would need a technologically feasible and impactful project.
After obtaining replicates of our results, he advised us to reach out to funding institutions Canadian Institutes of Health Research and produce scientifically sound data.
Then, we would work on the implementation and scaling up of the project before reaching out to humanitarian organizations and larger international funding
institutes for distribution of our product.
Andrew is the team’s expert in protein modelling.
His deadpan, straight-faced humour lightens up everyone’s mood even on the most stressful days.
He keeps the team morale up with his cheery attitude and off-the-charts optimism.
He speaks English, ASL, and just enough French to order a bagel.
In his free time, he is a runner, swimmer, and an equestrian.
Andrew is the team’s expert in protein modelling.
His deadpan, straight-faced humour lightens up everyone’s mood even on the most stressful days.
He keeps the team morale up with his cheery attitude and off-the-charts optimism.
He speaks English, ASL, and just enough French to order a bagel.
In his free time, he is a runner, swimmer, and an equestrian.
Andrew is the team’s expert in protein modelling.
His deadpan, straight-faced humour lightens up everyone’s mood even on the most stressful days.
He keeps the team morale up with his cheery attitude and off-the-charts optimism.
He speaks English, ASL, and just enough French to order a bagel.
In his free time, he is a runner, swimmer, and an equestrian.
The feedback we received from our stakeholders was that a bioreactor capable of producing a steady supply of vitamin-A producing yeast would be much better than waiting for biannual distribution of the vitamin. Omega-3 fatty acids are an emerging essential nutrient with numerous health benefits which make its production favourable.
Dr. Sanou mentioned that even with omega-3 supplementation, intestinal parasites will still pose a problem in preventing the absorption of not only vitamin A but the omega-3 fatty acids too. Because of this, our team decided to change the production of omega-3 to that of thymol, an anthelmintic compound found in thyme leaves. The compound is shown to critically damage Caenorhabditis elegans and its eggs, which is the gold standard in anthelmintic drug testing. While this is not a cure to intestinal parasites, it can mitigate intestinal damage and improve general micronutrient absorption until drugs such as Albendazole are distributed, even without access to clean water or proper footwear.
In places like the Philippines, India, Burkina Faso, and Uganda, people would have events like “nutrition week” or “micronutrient week” dedicated to diagnosing VAD, deworming using Albendazole, supplying vitamin A, and also informing parents and children about the importance of incorporating vitamin A in the diet. Dharamwati thinks that having a communal bioreactor would be better than individual ones for homes as it would be built with shared investment, and encourage community participation especially in events such as nutrition week.
We talked to Tyler Warnock, a medical school student who has worked with Healthy Child Uganda and Mama na Mtoto in Tanzania about his experience with global health work in Uganda, and how we could apply them towards our bioreactor and project implementation. He cautioned that making a difference is all about relationship building, and its absence would make our project implementation very difficult and “ unethical”. Many western people come in to collect data and leave without infrastructure, causing a divide between them and the local people.
With unethical implementation and inaccurate testing in mind, our team worked on the Randle Cell Testing Device which detects vitamin A levels in the blood. This is important because it helps us build a more accurate picture of VAD in our target areas, and informs us and our end-users whether our solution is making a difference.
To learn more about the Randle Cell Testing Device and Thymol Production, please click the buttons below.
We also talked to Dr. Samir Gupta, one of the co-founders of the Health Program at the Global Pathway School in India, which was developed by Canadians to help Indians students living below the poverty-line. We learned about their application of a 15-point plan which includes feeding the students meals in school, and supplying vitamin A and Albendazole for deworming. We asked him how he and his team successfully built this community without being culturally insensitive. He said that it is important to employ local people in leadership roles and in positions which enable community members to connect with them around economic and social problems. For the implementation of our yeast, he suggested considering schools for distribution since kids would be in one location being fed.
Our HP contacts agree that having microenterprises linked to Oviita would make our project more self-sustaining as it would train the local people while providing empowerment and employment, especially to women. Taking Dr. Gupta’s feedback in consideration, we partnered with the West Nile Youth Empowerment Centre (WNYEC) in Uganda which empowers youth through cultural and entrepreneurial training across eight different regions in Uganda. After explaining our project and its microenterprise opportunities, Nadia Iddi, the programs officer of WNYEC, said that this would be a great idea as it would create employment opportunities in their area. As long as our team provided them the initial training, they would also train their community members in how to operate our bioreactor, creating products like yoghurt fortified with Oviita, and educate people with the importance of vitamin A in the diet. Once the proposed training model is set in place, our partnership would expand to nearby health clinics around the area. Nadia informed us that the general perception for genetically modified organisms in Uganda is negative, but that should not be a threat with its monetary and health benefits. Over the next year, we will be working closely with them to see how we could best integrate the Oviita system tailored to their resources and culture.
The graphic above represents the Oviita system we've build hand-in-hand (virtually) with our end-users after various iterations. Now that we had a stakeholder-approved proposal, it was time to figure out how to make the science work.
DESIGN SOLUTIONS
Expert Consultations
CELLULASE ENGINEERING
Vitamin A supplements are typically provided or sold by foreign bodies. If the supply-chain were disrupted, VAD-struck countries would not receive the vitamin. Unfortunately, this is the case with many humanitarian organizations such as Save The Children International, which halted supplement and material donations to vulnerable countries due to COVID-19. Thus, we needed to come up with a self-sustaining solution that the community could maintain. Many of these communities grow crops for a living, so we thought to engineer cellulases into our yeast. Since our team has no experience working with Yarrowia lipolytica, we needed to know what considerations were required for our system.
Lourlin Ugdiman
Public Health & Nutrition
Lourlin is a Public Health & Nutrition Worker working for the government in Arayat, Philippines. It was crucial for us to understand the status of Vitamin A deficiency in the Philippines, which was once a hotspot for VAD-caused child blindness. Lourlin also provided us insights on how supplements from the government reach the community.
Vitamin A is supplied every 6 months in the community every 6 months. The government provides the supplements while healthcare and nutrition workers like Lourlin provide them in schools, municipal centers, clinics, and individual houses that have difficulty going to these centers. The supplements they provide are sourced externally, such as India and China.
“Nagbibigay kami sa community ng vitamin A every 6 months. Ang pagbibigay namin ay tuwing April at October. Ayan (letrato) ang bigay ng Rural Health Unit sa DOH (Department of Health). Yung 200,000 IU binibigay namin sa mga batang 12 months to 59 months. Ang 100,000 IU ay kulay blue ay pang 9 months to 11 months. Hindi na katulad ng dati kasi now nasusuplayan ng government yung kailangan ng mga bata nga ng mga suppliment. Pero di nagiging sapat kasi sa kakulangan ng pagkain sa kani-kanilang pamilya. Kung may budget, gamit, at may personnel, para sa pag-gawa ng vitamin A, bakit hindi? Willing makipagtulungan ang gobyerno para sa ikauunlad at kapakanan ng kalusugan ng mga tao sa kumunidad.”
In our interview, Lourlin says “We supply vitamin A every 6 months at April and October. That (picture) is given by the Rural Health Unit in the Department of Health. The 200,000 IU we give to children 12-59 months, while the 100,000 IU which is blue (bottle) we give to kids 9 to 11 months. It’s not (VAD cases) like before because now the government provides the supplements. (As for the bioreactor) If there are finances, resources, and personnel, why not? The government is willing to work together as long as it’s for the development and welfare of the people and community’s health.”
Dr. Charles Mather
Dry Lab
Dr. Charles Mather is a professor of social and cultural anthropology, sub-Saharan anthropology, and medical anthropology at the University of Calgary. As an individual with considerable experience in sub-Saharan cultures and medical systems, we contacted Dr. Mather for insight into how sub-Saharan regions of Africa are affected by Vitamin A deficiency, and how they may go about implementing our solution.
After expressing his support of and enthusiasm for our idea, Dr. Mather provided us with valuable feedback that will no doubt inform our design process. For instance, he informed us that some rural areas lack access to electricity, which poses a challenge to the design of our bioreactor, into which we were considering integrating an electric air pump. He cited solar-powered air pumps as a feasible alternative, while also suggesting that the culturing process be modified to incorporate stirring, rather than needing an air pump. As extreme heat decreases the efficiency of the yeast, Dr. Mather also cautioned us of the climate in Ghana, which can reach over 50°C in the dry season.
Furthermore, Dr. Mather informed us that people in Northern Ghana are very willing to take initiative with regards to technologies that will have a positive impact, suggesting that a representative travel to the community to educate people about how to culture, harvest, and cook the yeast, as well as the health benefits of the yeast. Considering the fact that Northern Ghana has an active beer brewing industry, where local women take turns brewing beer in an outdoor setup for the entire community. Dr. Mather recommended adapting the yeast culturing and harvesting process so that it can be integrated into existing beer brewing infrastructure. He emphasized that in many cases it would not be important to tell the people exactly how to assemble or culture the yeast. Instead, in many respects it might be preferable to simply tell them the growing needs of the yeast and allow them to decide how best to meet those needs.
Dr. Mather also provided some insight into the cultural associations surrounding blindness, stating that spiritual healing medicines and shrines play a big role in Ghanaian culture. However, he cited the Ghanaian people as highly pluralistic and open-minded, stating that they are willing to adopt new beneficial technologies and medicines as they become available. In order to implement our solution in a long-term and sustainable manner, he recommends beginning at the village level. Once our technology has been adopted by the village, he then recommends expanding into surrounding areas and moving up the chain of command.
Dr. Sanou is a public health nutritionist who has worked with UNICEF, National Public Health Laboratory, and National Nutrition Center in Burkina Faso.
He received a Ph.D. in Nutrition from Laval University and completed his postdoctoral training in aboriginal health at the University of Alberta.
A graduate from the University of Ouagadougou (B.Sc. and M.Sc.), Professor Sanou also holds a Certificate in information technology and nutrition from
the Swedish University of Agricultural Sciences.
The goal of our meeting to understand what bigger organizations such as UNICEF do to fight VAD.
We also wanted to talk to him more specifically about the steps we would need to take from creating a product in the lab to its deployment in communities.
He gave us insights about the vitamin A fiasco and the reasons why projects such as golden rice and other biofortified crops don’t work.
This includes farmers preferring earning money for planting exportable crops over biofortified ones.
Additionally, carotenoid to retinol bioconversion in the human body is minimal since the crops do not have a steady supply and take long to grow.
With biofortification still being a new technology and thus making a very small part of the diet, Dr. Sanou says that having a regular supply of vitamin A
rather than twice a year would be “much better”.
He works with communities to have “micronutrient week” or “vitamin A week” where healthcare workers mass supply vitamin A and educate them about its importance in diet.
His concern though, is that even with more supplements sufficient for more than 100% of the country, there will still be individuals who will never access them due to
issues such as distance and lack of proper roads and infrastructures.
Dr. Sanou explained that moving to a larger scale for supplementation, we would need a technologically feasible and impactful project.
After obtaining replicates of our results, he advised us to reach out to funding institutions Canadian Institutes of Health Research and produce scientifically sound data.
Then, we would work on the implementation and scaling up of the project before reaching out to humanitarian organizations and larger international funding
institutes for distribution of our product.
Andrew is the team’s expert in protein modelling.
His deadpan, straight-faced humour lightens up everyone’s mood even on the most stressful days.
He keeps the team morale up with his cheery attitude and off-the-charts optimism.
He speaks English, ASL, and just enough French to order a bagel.
In his free time, he is a runner, swimmer, and an equestrian.
THYMOL PRODUCTION
Vitamin A supplementation is coupled with deworming with compounds like Albendazole twice a year. This is because intestinal parasites perforate the intestines and prevent the maximal absorption of vitamin A and other micronutrients. Despite biannual deworming, lack of access to clean water and proper footwear only causes intestinal parasites to return. Additionally, these worms may also develop drug resistance especially since anthelmintic and deworming agents commercially available are limited. To alleviate intestinal damage, we decided to genetically engineer our yeast to produce thymol, a compound found in thyme leaves. Thymol has been shown to have anthelmintic properties against Caenorhabditis elegans and their eggs.
Our team had no clue how anthelmintics worked, or how drugs like Albendazole transition from the laboratory to schools. To find out more, we talked to a molecular and developmental geneticist, a doctor, and a parasitologist.
Paul E. Mains
Professor of Biochemistry & Molecular Biology
Dr. Mains is a biochemistry and molecular biology professor at the University of Calgary. He works with the resistance of worms to the anthelmintic benzimidazole, and developing Caenorhabditis elegans embryos. His background was vital for planning the experimental design of thymol production.
Dr. Mains said there was merit to producing thymol in our yeast because 1-2 billion people are infected by nematodes, mainly school-age children. There are effective treatments, but based upon his experience mainly with sheep parasites, resistance is inevitable so new treatments should be kept on the radar.
Dr. Mains thought C. elegans might not be able to consume the yeast because of its thick cell wall, and advised us to perform a simple experiment where we place non-engineered yeast on a petri-dish with worms first and see if they an use the yeast as an energy source. Additionally, he said we shouldn’t place c. elegans on a yeast plate but put yeast in C. elegans plates instead since cholesterol is needed for their growth. Alternatively, we could also combine cultures of our modified yeast with wild-type Escherichia coli if thymol is excreted, or add thymol to E. coli and feed that to the worms.
We thank Dr. Paul for generously offering to pick worms and perform thymol tests on C. elegans to save our team some time from training, since worm mating and picking takes time to perfect.
Dr. Samir Gupta
Respiratologist & GPS Health Program Co-founder
Dr. Gupta is a Respirologist in the University of Toronto and is one of the co-founders of the Health Program at the Global Pathway School in India, which was developed by Canadians to help Indian students living below the poverty-line. His background as a medical doctor and a leader in developing the health program in the school was important for us learning about implementation and working in international communities.
We learned about their application of a 15-point plan which includes providing resources from workshops on life skills, immunization, to counseling parents and children. We were most interested in their feeding the students meals in school, and supplying vitamin A and Albendazole for deworming as it relates directly to our project.
Dr. Gupta raised concerns around the safety and efficacy of thymol compared to existing drugs like Albendazole. While we might think that having thymol would be more beneficial than waiting for biannual deworming, we should still prioritize thinking about its safety.
We asked Dr. Gupta what kinds of regulations we might need to consider if we were to deploy thymol from the laboratory to an edible supplement. We found out that pharmaceutical trials and supplement trials are done differently. Depending on the character of thymol, we might be able to market our yeast as a supplement rather than a pharmaceutical. This would be faster and more cost-effective than pharmaceutical trials which involve larger human trials. He also agreed that probiotics might be a good parallel for regulation of our product, but its genetic modification might change the regulatory system for Oviita.
Dr. gilleard also helped micha
BIOCONTAINMENT MEASURES
A synthetic biology project without a biocontainment strategy would simply be irresponsible. If we wanted our yeast to be deployed outside a lab setting and grown in community bioreactors, we needed to observe extra precautions, making sure we don't do ecological damage to nature. We initially played around with the idea of having a light-inducible kill-switch system, but we weren't sure if that was sufficient.
Thus, we talked to a series of biochemists and molecular biologists who knew a thing or two about effective biocontainment strategies.
Dr. Marija Drikic
Information Here
Dr. Gordon Chua
Information here
Dr. Vanina Zaremberg
Information here
Dr. Robert Mayall
Fredsense Industries
Dr. Peter J. Facchini
Information Here
BIOREACTOR DESIGN
Growing conditions for our yeast must be regulated so we can maintain sufficient beta-carotene production and control beta-carotene doses for consumption. Proper design is also needed to make sure our yeast is contained in a closed-system, preventing accidental ecological damage.
RANDLE CELL TESTING DEVICE
Available data on VAD is obsolete, dating back to 2005. This data was not obtained through testing of retinol concentrations in the blood, but through a series of diagnostic tests scrutinizing vision, height, and academic performance. For Oviita to really make a difference, we need to know whether our supplementation project makes a difference to our end-users. This is why we created the Randle Cell Testing Device, a field-based test which utilizes aptamers to quantify retinol binding protein concentrations in the blood. These concentrations can then be correlated to Vitamin A concentrations, enabling communities to detect VAD effectively.
We reached out to professionals from various engineering principles to help the device come to fruition.
Dr. Christopher Naugler
Information Here
Dr. Colin Dalton
Information here
Sultan Khetani
Information here
Dr. Robert Mayall
Fredsense Industries
Banda Ndiaye
Information Here
Evaluate & Iterate on Design
Further Improving Our Design
At iGEM Calgary, we are dedicated to see proposals come to fruition. For Oviita to truly make a difference, we have to consider all feedback with open arms. Even if this meant being roasted at various events.
FACULTY TALK
Shortly after forming our initial proposal, we assembled stakeholders ranging from nutritional anthropologists to electrochemists to evaluate our project proposal. Overall, the feedback we received was promising. However, many questions were raised towards the effectiveness of our light-inducible kill switch, our choice of chassis (Rhodotorula toruloides) and its safety status for consumption, and how we would assemble our constructs for engineering. We were informed that most kill switch systems are susceptible to mutations as there is little reason for the cell to keep the trait. Our system would also have problems if our yeast reaches open waters or goes deep into the soil where light levels vary from minimal to none. We were advised to explore auxotrophy or quorum sensing as an alternative biocontainment strategy. Many of our guests have not heard of our chassis before and expressed concerns about our ability to engineer it, let alone consume it. We were advised to consider this thoroughly, and use a food-safe yeast such as baker’s yeast instead. While we did not use baker’s yeast, we made the transition to Yarrowia lipolytica, a Generally Recognized As Safe (GRAS)-designated yeast which is oleaginous. Having an oleaginous yeast would aid in vitamin A delivery as beta carotene and retinol are lipophilic. The strain we graciously received from Dr. Amaro’s laboratory in London has been engineered to produce the carotenoid already. Lastly, our attendees educated us about the wonders of Gibson Assembly and how it would save us a lot of trouble while assembling the constructs we needed.
Julygem
We hosted a two-day online meetup in July, where teams from Canada, USA, Taiwan, and Mexico got the chance to share their projects and receive feedback from experienced judges, as well as hear a number of talks by experts in the field that followed the overall theme of translating synthetic biology into the real world. Teams were asked to prepare a 5-7 minute pitch on their project to be presented in front of judges that were not all experienced biologists. Teams that evaluated our pitch though that we should talk to more stakeholders and integrate their needs and feedback to our presentation. The teams also thought we could do a better job of explaining how our solution provides a competitive edge towards existing solutions in vitamin A supplementation, and how both wet lab and dry lab are working together to address the problem more clearly.
iGEM Concordia's Mini Jamboree
We were fortunate to be invited by iGEM Concordia to participate in the Mini Jamboree where we had a friendly competition with four other Canadian teams. The judges thought we were on the right track on working with the local taste palettes and engineering principles. We were cautioned to check our assumptions about the societies we were working with in terms of building materials. Our audience was a bit lost with the values for vitamin A and EPA and how they are linked together, and thought a model on our co-culture switch system would be worth considering. The other teams also agreed that we needed to provide more reasoning why our project was important.
cGEM
We participated at cGEM, a Canada-wide iGEM competition where we presented a poster and a 20-minute project presentation. The area where our judges and fellow teams thought we were lacking the most was showing the integration of our stakeholders throughout the process in our presentation, and whether we have talked to our end-users in vitamin-A deficient areas. At the time, we did not talk to local communities and public health workers so we made sure to do that and reached out to people in India, Philippines, and Uganda. In particular, they were concerned about having a training program or system set in place to ensure the communities knew how our bioreactor would operate, and how the yeast would be properly incorporated into their diets. We were also advised to address any possible obstacles we might face with our solutions